An RND-Type Efflux System in Borrelia burgdorferi Is Involved in Virulence and Resistance to Antimicrobial Compounds

Abstract

Borrelia burgdorferi is remarkable for its ability to thrive in widely different environments due to its ability to infect various organisms. In comparison to enteric Gram-negative bacteria, these spirochetes have only a few transmembrane proteins some of which are thought to play a role in solute and nutrient uptake and excretion of toxic substances. Here, we have identified an outer membrane protein, BesC, which is part of a putative export system comprising the components BesA, BesB and BesC. We show that BesC, a TolC homolog, forms channels in planar lipid bilayers and is involved in antibiotic resistance. A besC knockout was unable to establish infection in mice, signifying the importance of this outer membrane channel in the mammalian host. The biophysical properties of BesC could be explained by a model based on the channel-tunnel structure. We have also generated a structural model of the efflux apparatus showing the putative spatial orientation of BesC with respect to the AcrAB homologs BesAB. We believe that our findings will be helpful in unraveling the pathogenic mechanisms of borreliae as well as in developing novel therapeutic agents aiming to block the function of this secretion apparatus.

Author Summary

Lyme disease is caused by infection with the spirochete Borrelia burgdorferi. These spirochetes cycle between Ixodes ticks and vertebrate reservoirs, mainly rodents, but also birds. Previous studies have revealed major differences in the B. burgdorferi cell envelope structure and membrane composition compared to those of other bacteria. Proteins embedded in the bacterial membranes fulfill a number of tasks that are crucial for bacterial cells, such as solute and protein transport, as well as signal transduction, and interaction with other cells. Microorganisms have evolved mechanisms to protect themselves against harmful substances and secrete these through efflux pumps. So far, little is known about mechanisms of drug efflux systems in borreliae. Herein we identified an outer membrane channel forming protein important for B. burgdorferi to cause infection in mice and that also is involved in antibiotic resistance. We believe that this work will be helpful to understand the mechanisms underlying borreliae infection biology as well as in developing new therapeutic agents aiming to block this multi drug efflux system.